Neuron Specific Enolase (NSE) Antibody Staining Protocol for Immunohistochemistry

 

Description: Enolases are homo- or heterodimers of the three subunits: alpha (46kDa), beta (44kDa), and gamma (46kDa). The alpha-subunit is expressed in most tissues and the beta-subunit only in muscle. The gamma-subunit is expressed primarily in neurons, in normal and in neoplastic neuroendocrine cells. Coexpression of NSE and chromogranin A is common in neuroendocrine neoplasms. Gamma-Gamma isozyme of enolase is found at elevated concentrations in small cell lung cancer and pediatric neuroblastoma.

 

Primary Antibody

Name: Neuron Specific Enolase (NSE) Antibody

Clone: Rabbit anti-Human

Supplier: Dako

Catalog Number: A0587

Dilution: 1:2000 using IHC-TekTM Antibody Diluent (Cat# IW-1000 or IW-1001) to reduce background and unspecific staining and serum blocking step is NOT needed.

Incubation Time/Temp: 60 min/room temperature


Antigen Retrieval
Device: IHC-TekTM Epitope Retrieval Steamer Set (Cat# IW-1102)
Buffer/pH value: IHC-TekTM Epitope Retrieval Solution (Cat# IW-1100)
Heat/Cool Temperature: 95-100 ºC/room temperature
Heat/Cool Time: 20 minutes/20 minutes

Detection Methods
Standard Method: ABC Method or LSAB Method
Enhanced Method: Polymeric Methods

Chromogen Substrate
Reagent: DAB
Incubation Time/Temperature: 1-3 minutes/room temperature

Counterstain
Reagent: Mayer's Hematoxylin
Staining Time: 30 seconds

Results:
Staining Pattern: Cytoplasmic
Images: Search image

Additional Information:
Species Reactivity: Human
Fixation: Formalin fixed paraffin sections
Positive Control: Brain
Negative Control: Omit primary antibody, isotype control, absorption control
Blocking: 2-5% normal serum to reduce unspecific background staining; 0.5-3% H2O2 to block endogenous peroxidase activity; avidin/biotin to block endogenous biotin activity if necessary

 

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